Theriogenology最新文献

{"title":"The analysis of X chromosome activity of porcine embryonic stem Cells: Study based on parthenogenetic embryonic stem cells with LCDM medium","authors":"Yu Shi ,&nbsp;Hongxing Wang ,&nbsp;Mengjia Chai ,&nbsp;Mengru Ji ,&nbsp;Wenqian Zhao ,&nbsp;Qianqian Xu ,&nbsp;Tingsheng Yan ,&nbsp;Zhonghua Liu ,&nbsp;Xiaogang Weng","doi":"10.1016/j.theriogenology.2025.117479","DOIUrl":"10.1016/j.theriogenology.2025.117479","url":null,"abstract":"<div><div>The derivation of porcine embryonic stem cell (pESC) lines remains a major challenge in this field. To date, the porcine naïve ESCs have yet to be successfully established, and standardized criteria for their characterization and evaluation are still lacking. The regulation of X-chromosome activity integrates information from embryonic development and the dosage of sex chromosomes, which is closely associated with the pluripotent state of embryonic stem cells. In this study, we aimed to establish pESC lines in LCDM medium from porcine blastocyst-stage embryos, and analyzed the features of ESCs from the sight of X chromosome activity. We assessed molecular markers and epigenetic characteristics to confirm pluripotency and X chromosome activity in porcine parthenogenetic ESCs (named as ppLCDM) using <em>XIST</em> RNA-FISH, immunofluorescence staining, single-cell RNA sequencing (scRNA-seq), and other techniques. Results showed that ppLCDM cells expressed most pluripotent markers. The percentage of ppLCDM cells exhibiting H3K27me3 and <em>XIST</em> aggregation signals increased with passage, indicating the progressive establishment of X-chromosome inactivation (XCI). Meanwhile, the pluripotency of most ppLCDM cells gradually declined during extended passaging. However, two distinct patterns of ppLCDM cells were observed from passage 35 (type I cells, P35-I) displayed normal XCI states, while type II cells (P35-II) exhibited X-chromosome erosion-like state, characterized by the loss of aggregation signals, abnormal X-linked gene ratios. Particularly, the pluripotency of ppLCDM cells with an X-chromosome erosion-like state undergoes unusual changes compared to normal cells. These findings indicate that X chromosome activity is closely associated with the pluripotent state of porcine ESCs and that heterogeneity in X chromosome activity arises during passaging. Our research provides crucial insights into X chromosome dynamics in large-animal ESC models and contribute to ongoing efforts to establish stable naïve pESC lines.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117479"},"PeriodicalIF":2.4,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143936778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing embryonic and full-term development during mouse cloning through post-activation treatment with JNJ-7706621","authors":"Hyo-Gu Kang ,&nbsp;Min Ju Kim ,&nbsp;Ji Hyeon Yun ,&nbsp;Eun Young Choi ,&nbsp;Se-Been Jeon ,&nbsp;Pil-Soo Jeong ,&nbsp;Bong-Seok Song ,&nbsp;Ki-Hoan Nam ,&nbsp;Sun-Uk Kim ,&nbsp;Min Kyu Kim ,&nbsp;Bo-Woong Sim","doi":"10.1016/j.theriogenology.2025.117475","DOIUrl":"10.1016/j.theriogenology.2025.117475","url":null,"abstract":"<div><div>Somatic cell nuclear transfer (SCNT) is widely researched for animal cloning. However, SCNT embryos frequently exhibit reduced developmental potential compared to those generated through natural reproduction. This study aimed to improve SCNT mouse embryo development by assessing the effects of JNJ-7706621 (JNJ, a specific inhibitor of cyclin-dependent kinase 1 and aurora kinases) as a post-activation treatment, replacing cytochalasin B (CB). Parthenogenetically activated (PA) mouse embryos were cultured with CB (5 μg/mL) or JNJ (1, 10, or 50 μM) to determine the optimal concentration. The 10 μM JNJ and CB groups showed significantly higher developmental competency compared to the 1 and 50 μM JNJ groups. The 10 μM JNJ group also exhibited an increase in total cell number and a decrease in apoptotic cells compared to the CB group. SCNT mouse embryos treated with 10 μM JNJ showed improved development (CB: 39.9 % ± 6.4; JNJ: 61.4 % ± 4.4), with increases in total cell number (CB: 52.7 ± 3.6; JNJ: 70.7 ± 2.9), inner cell mass (CB: 10.4 ± 0.7; JNJ: 15.4 ± 1.1), and trophectoderm cells (CB: 42.3 ± 3.3; JNJ: 55.3 ± 2.5). JNJ treatment significantly reduced aberrant F-actin and tubulin compared to CB. It also reduced abnormal spindles in one-cell embryos and decreased blastomere fragmentation and DNA damage in two-cell SCNT embryos compared to CB. Importantly, JNJ treatment led to significantly higher implantation (CB: 50.8 % ± 3.7; JNJ: 68.3 % ± 4.3) and live birth rates (CB: 2.4 % ± 2.4; JNJ: 10.9 % ± 2.8) compared to CB. These results demonstrate that JNJ enhances cytoskeletal integrity and chromosome stability, ultimately improving both preimplantation development and full-term success in mouse SCNT embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117475"},"PeriodicalIF":2.4,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143936777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of Na+⁄K+-ATPase associated with boar fertility","authors":"Muhammad Imran ,&nbsp;Murray Pettitt ,&nbsp;Mary M. Buhr","doi":"10.1016/j.theriogenology.2025.117460","DOIUrl":"10.1016/j.theriogenology.2025.117460","url":null,"abstract":"<div><div>The ubiquitous transmembrane protein Na⁺⁄K⁺-ATPase affects sperm fertility and capacitation through ion transport and cell signaling, but its effect on <em>in vivo</em> boar fertility, and any controlling mechanism(s) are unknown. The isoforms of the α and β subunits of Na⁺⁄K⁺-ATPase (α1, α2, α3, β1, β2, β3) were assessed in one ejaculate from each of six high and six low fertility boars (HF, LF respectively) which differed in their <em>in</em> vivo direct boar effects (DBE) for both farrowing rate (FR) and litter size (p &lt; 0.05). Sperm function assessed by CASA was similar in HF and LF ejaculated sperm. Immunocytochemistry of the ejaculate sperm found more intact HF than LF sperm had α3 equatorially (p &lt; 0.05), and fewer HF than LF methanol-permeabilized sperm had detectable α2 (p &lt; 0.05). Image Quant analysis of Western blots rigorously quantified the amount of each isoform in the isolated head plasma membrane from each ejaculate, determining that the total amount of the isoform α3, and the amounts of 7 individual α3 bands, differed significantly (HF &gt; LF, p &lt; 0.05), and linear regression confirmed a highly significant relationship of total amount of α3 and α1 with FR (r² = 0.90; p &lt; 0.0001 and r² = 0.36; p &lt; 0.05, respectively). The β1, β2 and β3 isoforms in LF lacked several molecular weight bands common to HF, and the total volume of β1 tended to correlate with DBE for FR (r² = 0.2652; p = 0.09). These findings suggest that specific isoforms of Na⁺/K⁺-ATPase in the sperm head are correlated to boar <em>in vivo</em> fertility, probably through Na⁺⁄K⁺-ATPase's role in capacitation.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117460"},"PeriodicalIF":2.4,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143947490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect of cryopreservation on the turkey (Meleagris gallopavo) semen proteome","authors":"Laura Pardyak ,&nbsp;Ewa Liszewska ,&nbsp;Sylwia Judycka ,&nbsp;Sylwia Machcińska-Zielińska ,&nbsp;Halina Karol ,&nbsp;Mariola A. Dietrich ,&nbsp;Ewa Gojło ,&nbsp;Zbigniew Arent ,&nbsp;Barbara Bilińska ,&nbsp;Giusy Rusco ,&nbsp;Nicolaia Iaffaldano ,&nbsp;Andrzej Ciereszko ,&nbsp;Mariola Słowińska","doi":"10.1016/j.theriogenology.2025.117473","DOIUrl":"10.1016/j.theriogenology.2025.117473","url":null,"abstract":"<div><div>Two-dimensional difference in-gel electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry was employed to investigate the proteome alterations induced by equilibration and freezing/thawing processes, both in turkey spermatozoa and in extracellular fluid (ECF). The freezing/thawing process resulted in reduced semen quality parameters. Viability and motility decreased threefold (90.6 %–31.2 % and 76.0 %–26.7 %, respectively), while the proportion of live spermatozoa with intact mitochondrial membrane potential decreased fivefold (54.9 %–11.4 %). Additionally, oxidative stress increased sevenfold (10.5 %–68.8 %). A total of 146 differentially abundant protein spots were found between fresh and frozen/thawed spermatozoa, while 27 spots differentiated between fresh and frozen/thawed ECF. Immunofluorescence staining showed reduced signals of mitochondrial proteins, such as aconitate hydratase, alpha-enolase, glycerol-3-phosphate dehydrogenase, and triosephosphate isomerase, in the spermatozoa midpiece, as well as reduced signals of acrosin in the acrosome. Freezing/thawing affected mitochondrial energy metabolism, particularly the tricarboxylic acid (TCA) cycle and oxidative phosphorylation. The maintenance of an acetyl-CoA pool to sustain TCA cycle activity in cryopreserved spermatozoa may be insufficient owing to disturbances in fatty acid beta-oxidation and/or aerobic glycolysis. Changes in the ECF primarily reflect the leakage of spermatozoa glycolytic enzymes. The freezing/thawing process alters motile cilium assembly, primarily affecting the spermatozoa axoneme and outer dense fibres. The initial step of fertilization may be disrupted by alterations in proteins involved in spermatozoa binding to the ovum. These findings extend our knowledge of the molecular and cellular mechanisms of cryodamage in turkey semen which is prerequisite for the improvement of semen preservation procedures.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117473"},"PeriodicalIF":2.4,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143934921","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Supplementation with lipoamide during in vitro maturation improves bovine oocyte maturation and subsequent embryonic development: potential link to PI3K/AKT signaling","authors":"Canqiang Lu ,&nbsp;Ziyun Ruan ,&nbsp;Yun Wang ,&nbsp;Zhenhua Tang ,&nbsp;Dongping Zhou ,&nbsp;Yiqing Yang ,&nbsp;Yanyu Chen ,&nbsp;Jie Ren ,&nbsp;Chengxi Zeng ,&nbsp;Zhengda Li ,&nbsp;Deshun Shi ,&nbsp;Fenghua Lu","doi":"10.1016/j.theriogenology.2025.117417","DOIUrl":"10.1016/j.theriogenology.2025.117417","url":null,"abstract":"<div><div>Oxidative stress during oocyte <em>in vitro</em> maturation (IVM) is still concerned. Lipoamide (LAM) has been widely studied as an agent for alleviating various diseases associated with oxidative disruption. This work aimed to evaluate the potential effects of LAM on bovine oocyte IVM and its mechanisms. Different concentrations of LAM (0, 10, 50, 100, and 200 μmol/L) were supplemented to bovine oocyte IVM medium. The IVF derived zygote cleavage and blastocyst formation rate in the 100 μmol/L LAM treatment group was increased compared with the control group (<em>P</em> &lt; 0.05).There was no statistical difference in PBI between 100 μmol/L LAM treatment and the control group, although the treatment tended to increase it (<em>P</em> = 0.059). Further revealed that LAM increased the expression of PI3K and phosphorylated-AKT1 (pAKT1), improved mitochondrial profile, and reduced apoptosis in bovine oocytes. Meanwhile, the reactive oxygen species (ROS) as well as the 8-Hydroxydeoxyguanosine (8-OHdG, DNA damage-specific marker) displayed lower levels accumulation in LAM-exposed oocytes. Taken together, the results show that administration of LAM (100 μmol/L) during IVM can ameliorate the developmental competence of bovine oocyte through the potential regulation of oxidative stress, apoptosis, DNA damage, and PI3K/AKT signaling.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117417"},"PeriodicalIF":2.4,"publicationDate":"2025-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143912148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"H-Y antigen-based immuno-segregation strategies for bovine X sperm enrichment","authors":"Richa Khirbat ,&nbsp;Trilok Nanda ,&nbsp;Aman Kumar ,&nbsp;Meenakshi Virmani ,&nbsp;Akhil Kumar Gupta ,&nbsp;Jatin ,&nbsp;Sushila Maan ,&nbsp;Vinay G. Joshi","doi":"10.1016/j.theriogenology.2025.117474","DOIUrl":"10.1016/j.theriogenology.2025.117474","url":null,"abstract":"<div><div>Insemination of sperm bearing X and Y chromosomes prior to sexual pre-selection significantly impacts the livestock industry economically. Fluorescence-activated cell sorting (FACS) is a widely accepted technology that sorts sperm with over 90 % accuracy. High costs and potential damage to sperm limit the application of flow cytometry. Consequently, flow cytometry has been replaced by less intrusive and more affordable immunological methods. The present study hypothesizes that antibodies raised against male-enhanced antigen-1 (MEA-1) peptide of <em>Bos indicus</em> will specifically bind to bovine Y sperm, thus assisting in the enrichment of X sperm. A solid surface strategy (SSS) was employed to enrich the X chromosome-bearing spermatozoa of <em>Bos indicus</em>, utilizing purified IgG anti-peptide (MEA P-1) antibodies alongside commercial Human MEA-1 antibodies for comparison. A quantitative SYBR green real-time PCR assay targeting the Y chromosome-specific <em>SRY</em> gene and the X chromosome-specific <em>PLP</em> gene was conducted to determine the ratio of X and Y sperm in the enriched semen samples. The solid surface strategy using commercially available Human MEA-1 antibodies and anti-MEA P-1 antibodies yielded 34.72 ± 6.74 (X: Y; 73:27) and 20.02 ± 4.08 (X: Y; 64:36) % of X sperm enrichment, respectively. The mean percentage of X chromosome content in an unsorted semen sample and cow bull blood was 50.53 ± 0.19 and 49.90 ± 1.27, respectively, aligning with an expected ratio of 1:1. The sperm viability analysis using computer-assisted semen analyzer (CASA) for motility parameters and the hypo-osmotic swelling (HOS) test showed acceptable sperm viability post-treatment with anti-MEA P-1 antibodies. The study evaluated the efficacy and utility of anti-peptide MEA P-1 antibodies as an alternate approach for bovine X sperm enrichment.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117474"},"PeriodicalIF":2.4,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143922646","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “A novel loop-mediated isothermal amplification approach for sex identification of Columbidae birds” [Theriogenology 78 (2012) 1329–1338]","authors":"Kun-Wei Chan ,&nbsp;Pan-Chen Liu ,&nbsp;Wei-Cheng Yang ,&nbsp;James Kuo ,&nbsp;Cicero Lee-Tian Chang ,&nbsp;Chi-Young Wang","doi":"10.1016/j.theriogenology.2025.117459","DOIUrl":"10.1016/j.theriogenology.2025.117459","url":null,"abstract":"","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117459"},"PeriodicalIF":2.4,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143899633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Standardization of bovine sperm concentration analysis in a multi-laboratory setting","authors":"Leonardo F.C. Brito,&nbsp;Clara González-Marín,&nbsp;Pablo J. Ross","doi":"10.1016/j.theriogenology.2025.117472","DOIUrl":"10.1016/j.theriogenology.2025.117472","url":null,"abstract":"<div><div>Sperm concentration evaluation is a fundamental component of semen analysis, as results are critical for clinical management and preparation of semen for assisted reproduction. However, significant variations in sperm concentration measurements have been reported within and across laboratories. The objective of this study was to assess the effectiveness of standardization of bull sperm concentration analysis in a multi-laboratory setting. In Experiment I, NucleoCounter calibration (27 units, seven laboratories) was evaluated using frozen reference standards analyzed weekly over 12 weeks by the same technician in each laboratory. NucleoCounter performance was satisfactory and the few data points flagged on Levey-Jennings charts seemed to result from systemic errors introduced by the technician. The intra-technician coefficient of variation (CV) was 2.2 ± 1.8 % (mean ± SD), while the intra-equipment CV was 4.0 ± 1.3 %. In Experiment II, the effect of customized e-learning training was evaluated across six laboratories. When compared to results obtained before training, CV for duplicate results (3.2 ± 3.8 %) and proportion of samples with duplicate results differing by more than 10 % (8.1 %) decreased (P &lt; 0.05) after training (3.0 ± 3.2 % and 6.9 %, respectively). In Experiment III, 10 batches of frozen semen were produced with three replicates of each batch coded differently. Sperm concentration was evaluated by nine technicians across six laboratories during three test periods over one year. Intra-technician and intra-batch CV were 3.4 ± 3.1 % and 4.6 ± 2.2 %, respectively. When Bland-Altman plots showed that the mean difference of individual results and the overall mean was close to zero during all test periods (-0.36 to 0.55 × 10⁶ sperm/mL for the 17 × 10⁶ sperm/mL standard and -2.2 to 3.5 × 10⁶ sperm/mL for the 150 × 10⁶ sperm/mL standard). In conclusion, precise and accurate sperm concentration results were obtained by qualified technicians using standardized procedures and the NucleoCounter.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117472"},"PeriodicalIF":2.4,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143907836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chromosome remodeling and cytoplasmic distribution during embryonic development in fused pair embryos","authors":"Aochen Yu,&nbsp;Yang Xing,&nbsp;Fuyao Zhang,&nbsp;Deru Cao,&nbsp;Yuan Jiang,&nbsp;Xinyue Lu,&nbsp;Juan Li","doi":"10.1016/j.theriogenology.2025.117465","DOIUrl":"10.1016/j.theriogenology.2025.117465","url":null,"abstract":"<div><div>Cell fusion is now widely employed as an <em>in vitro</em> model for inducing and investigating cell fate changes. In early embryo research, fusion of embryos is also a way to probe the mechanisms of mammalian oogenesis and preimplantation development. To establish a novel model for porcine early embryo studies and investigate its developmental mechanisms, pair of zona pellucida (ZP)-free oocytes were electro fused to produce the fused pair (FP) embryos, which were further <em>in vitro</em> cultured to the blastocyst stage. Firstly, developmental competence was assessed, revealing a cleavage rate of 92.27 ± 5.59 % and a blastocyst rate of 26.12 ± 6.61 %, which were similar to the parthenogenetic activation (PA) embryos (<em>p</em> &gt; 0.05). Subsequently, nuclear and spindle staining was performed on FP embryos collected at 14–22 h, with 67.18 ± 3.18 % of the embryos spindle reorganization occurred and nuclei fusion, whereas a few displayed independent division of the two nuclei or tripolar spindle. Lastly, the distribution of lipid droplets (LDs) and mitochondria in FP embryos was assessed via fluorescent staining. Results showed that the even distribution of LDs from one oocyte was observed in each blastomere of 4-cell to blastocysts. A similar distribution pattern was observed for mitochondria, which was being observed at the 2-cell stage, a relatively earlier developmental stage than that of LDs. Results suggested that cytoplasm including mitochondria and LDs could redistribute once two oocytes fused into a single embryo. More studies are needed for the underlying mechanism and potential impact on the developmental ability of FP embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117465"},"PeriodicalIF":2.4,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143899550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Colony-stimulating factor 2 (CSF2) does not significantly affect cellular and molecular parameters, or blastocyst rates under in vitro culture conditions with reduced nutrient concentrations","authors":"Ana Caroline Chaves Vall Nicolás ,&nbsp;Ligiane de Oliveira Leme ,&nbsp;Amanda Oliveira Moura ,&nbsp;Nayara Ribeiro Kussano ,&nbsp;Emivaldo de Siqueira ,&nbsp;Maurício Machaim Franco ,&nbsp;Margot Alves Nunes Dode","doi":"10.1016/j.theriogenology.2025.117464","DOIUrl":"10.1016/j.theriogenology.2025.117464","url":null,"abstract":"<div><div>Colony-stimulating factor 2 (CSF2) is an embryokine used between days 5 and 7 of culture to improve embryonic development. We evaluated its use from Day 4 and throughout the culture period in a reduced nutrient concentrations culture medium. Presumptive zygotes were assigned to three treatments: Control – synthetic oviduct fluid (SOF), CSF2 D1 (SOF + CSF2 from Day 1–7), CSF2 D4 (SOF + CSF2 from Day 4–7). Cleavage rates on Day 4 and blastocyst rates on days 6 and 7 were evaluated. On Day 7, expanded blastocysts were used to assess mitochondrial activity, lipid quantification, total cell number, gene expression and DNA methylation patterns. Embryo development data were analyzed using the chi-square test (P ≤ 0.05). Data normally distributed were analyzed with ANOVA and Tukey's test, while non-normally distributed data were evaluated using the Kruskal-Wallis and Mann-Whitney tests. Cleavage and blastocyst rates on Day 7 were similar (P &gt; 0.05) among treatments. Of 11 genes evaluated, only Placenta-specific 8 presented a lower number of transcripts (P ≤ 0.05) in embryos cultured with CSF2 from Day 4. Assessments of DNA methylation, lipid droplet staining, mitochondrial activity and total cell number showed no significant differences among treatments (P &gt; 0.05). In conclusion, CSF2 had no effect on the assessed parameters when a culture medium with reduced nutrient concentrations was used, and can be used throughout the entire culture period without affecting embryo production and developmental kinetics compared to CSF2 added on Day 4.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"243 ","pages":"Article 117464"},"PeriodicalIF":2.4,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143907835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}